Purification and characterization of a hydrolysis‐resistant lipase from Aspergillus terreus. (21st November 2013)

• Record Type: Journal Article
• Authors: Shi, Hui; Meng, Yao; Yang, Min; Zhang, Qinglian; Meng, Yanfa
• Is Part Of: Biotechnology and applied biochemistry. Volume 61:Number 2(2014)
• Page Start: 165
• Page End: 174
• Publication Date: 2013-11-21
• Subjects: Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1142
• Languages: English
• ISSNs: 0885-4513

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <p>Lipase from <italic>Aspergillus terreus</italic> was purified to homogeneity using ammonium sulfate precipitation and chromatographies with Q‐Sepharose and Sephacryl S‐200. It showed a single band on SDS‐PAGE and IEF‐PAGE with a relative molecular mass of 37.2 kDa and p<italic>I</italic> of 3.2. Its glycoprotein nature was confirmed with the percentage of saccharides of 5.02% and 3.88% determined by the phenol/sulfuric acid and anthrone/ sulfuric acid methods, respectively. The lipase hydrolyzed both plant oils and animal oils, with the <italic>K</italic>_m value for substrate <italic>p</italic>‐NPP of 16.42 mM at pH 6.0, 50 °C. The enzyme was tolerant in a wide range of pH (pH 3–12) with optimum activity at pH 4.0. It remained stable under the highest temperature of 65 °C, with maximal activity at 50 °C. Ca²⁺, Co²⁺, Mn²⁺, and Ni²⁺ stimulated enzyme activity, but Hg²⁺ caused inhibition. Detected detergents had no obvious effect on enzyme activity, except SDS, which stimulated the activity at lower concentrations but inhibited the activity at higher concentrations. The inhibitory effect on enzyme activity of phenylmethanesulfonyl fluoride revealed that the Ser was involved in catalysis. Saccharides had no obvious effect on enzyme activity but could enhance its thermostability. Furthermore, the enzyme was resistant to trypsin digestion.</p> </abstract>